Active Carbonic Anhydrase IV (CA4)

CA-IV; CAIV; Car4; RP17; Retinitis Pigmentosa 17(Autosomal Dominant); Carbonate dehydratase IV

ACTIVITY TEST

Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO2  H2O = HCO3-  H , which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption. CA4 is a GPIanchored membrane enzyme expressed on the luminal surfaces of pulmonary (and certain other) capillaries and of proximal renal tubules. It functions as the principal CO2 taste sensor. The activity of recombinant human CA4 was measured by its ability to hydrolyze 4-Nitrophenyl acetate (4-NPA) to 4-Nitrophenol. The reaction was performed in 12.5 mM Tris, 75 mM NaCl, pH 7.5 (assay buffer), initiated by addition 50 μL of various concentrations of CA4 (diluted by assay buffer) to 50 µL of 2 mM substrate 4-NPA (100 mM stock in Acetone, diluted by assay buffer). Incubated at 37℃ for 5min, then read at a wavelength of 400 nm.

One unit of enzyme activity is defined as the 1 μg of enzyme required to convert 1 pmol of 4-Nitrophenyl acetate to 4-Nitrophenol in 1min at 37°C. The specific activity of recombinant human CA4 is > 15 pmol/min/µg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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